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长链非编码 RNA OIP5-AS1 通过靶向 miR-369-3p 调控 DYRK1A 增强结直肠癌细胞的放射抵抗。

LncRNA OIP5-AS1 regulates radioresistance by targeting DYRK1A through miR-369-3p in colorectal cancer cells.

机构信息

Department of Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei, China.

Department of Biochemistry and Molecular Biology, School of Basic Medicine, Huazhong University of Science and Technology, Wuhan, 430030, Hubei, China.

出版信息

Eur J Cell Biol. 2018 Jun;97(5):369-378. doi: 10.1016/j.ejcb.2018.04.005. Epub 2018 Apr 14.

DOI:10.1016/j.ejcb.2018.04.005
PMID:29773344
Abstract

OBJECT

This study aimed to investigate the role of lncRNA OIP5-AS1 in regulating radioresistance of colorectal cancer (CRC) cells.

METHODS

Microarray analysis was used to screen out lncRNAs differentially expressed in radio-resistant CRC cell lines. Expression levels of OIP5-AS1, miR-369-3p and DYRK1A in CRC cell lines were measured by qRT-PCR. Protein expression of DYRK1A was determined by western blot. The target relationships among OIP5-AS1, miR-369-3p and DYRK1A were validated by dual luciferase reporter assay. Impacts of OIP5-AS1 or DYRK1A on CRC cellular activity and apoptosis were investigated by MTT assay, clonogenic survival assay and flow cytometry to analyze OIP5-AS1 or DYRK1A's effect on radioresistance of CRC cells.

RESULTS

LncRNA OIP5-AS1 and DYRK1A were down-regulated in radio-resistant CRC cell lines. OIP5-AS1 suppressed the expression of miR-369-3p, thus up-regulating DYRK1A, the downstream gene of miR-369-3p. OIP5-AS1 and DYRK1A impaired cell clonogenic survival and promoted cell apoptosis after irradiation, improving radiosensitivity of CRC cells.

CONCLUSION

LncRNA OIP5-AS1 suppressed cell viability, promoted radio-induced apoptosis, and enhanced the radiosensitivity of CRC cells by regulating DYRK1A expression through miR-369-3p.

摘要

目的

本研究旨在探讨长非编码 RNA OIP5-AS1 在调节结直肠癌（CRC）细胞放射抵抗中的作用。

方法

采用基因芯片分析筛选出耐辐射 CRC 细胞系中差异表达的 lncRNAs。采用 qRT-PCR 检测 CRC 细胞系中 OIP5-AS1、miR-369-3p 和 DYRK1A 的表达水平。采用 Western blot 检测 DYRK1A 蛋白表达。采用双荧光素酶报告基因实验验证 OIP5-AS1、miR-369-3p 和 DYRK1A 之间的靶标关系。通过 MTT 检测、集落形成实验和流式细胞术分析 OIP5-AS1 或 DYRK1A 对 CRC 细胞活性和凋亡的影响，以研究 OIP5-AS1 或 DYRK1A 对 CRC 细胞放射抵抗的影响。

结果

在耐辐射 CRC 细胞系中，lncRNA OIP5-AS1 和 DYRK1A 表达下调。OIP5-AS1 抑制 miR-369-3p 的表达，从而上调 miR-369-3p 的下游基因 DYRK1A。OIP5-AS1 和 DYRK1A 降低了细胞集落形成能力，促进了照射后的细胞凋亡，从而提高了 CRC 细胞的放射敏感性。

结论

通过 miR-369-3p 调控 DYRK1A 表达，lncRNA OIP5-AS1 抑制细胞活力，促进放射诱导的凋亡，增强 CRC 细胞的放射敏感性。

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